Identifying gut microbiota diversity in children born through normal delivery and cesarean section

IntroductionNewborns overall health and lifetime development are greatly influenced by their gut flora, which has an effect on their short- and long-term well-being. The immune system, mental health issues, and metabolic diseases have all been associated with abnormalities in gut microbiota, highlighting the critical role that gut microbiota plays in determining one’s quality of life. The gut microbiota functions as an extension of the host genome, containing 50-100 times more genes than the host and providing vital enzymes that the host is unable to manufacture. Many disorders can result from any changes in this microbial mix, known as dysbiosis.

The feeding schedule and mode of delivery are two examples of internal and external factors that have a significant impact on an infant’s gut microbiome health. Delivery methods such as vaginal and caesarean sections are crucial in determining the microbiological makeup. Because lactobacilli are so prevalent in the vagina, newborns delivered vaginally have higher than average quantities of these bacteria. On the other hand, babies born by caesarean section have lower microbe counts from the bacteroides genus.The composition of the gut microbiota is closely related to the immune system. Research has indicated that the gut microbiota plays a significant trole in controlling the immune system, and it has been linked to allergic and hypersensitive reactions via affecting the expression of the FOXP3 gene.So in this particular project we will be looking to decipher the effect of the mode of delivery on the gut microbiota diversity of the infant.

ObjectivesTo comprehend gut microbial diversity between neonates delivered vaginally and by caesarean section.

Study DesignA Prospective Cohort study.We will recruit healthy neonates delivered vaginally and by caesarean section at BLDE (DU)’s Shri B M Patil Medical Collage, Hospital & Research Centre, Vijayapura.During the first six months of their lives, we will monitor them and take stool sample three times: at birth (0 months), three months, and six months.

Study ProtocolFecal samples from infant delivered after the 36 weeks gestation (0 months), after 03 months and 06 months will be collected in sterile containers and transported to the research laboratory either frozen (-20°C home freezer) or fresh (if collected within 4 hours) and after aliquoting they will be stored at -80°C until analysis.Total DNA will be extracted using the Qiagen PowerSoil DNA Isolation Kit, Cat_12888100 and stored at -80°C until analysis.Universal primers for the V4 region of the16S IRNA gene will be used to amplify a 292 bp product that was sequenced on the Illumina MiSeq platform.USEARCH software will be used to cluster paired-end reads into OTUs. The mothur software suite will be used to assign representative sequences to taxa in the SILVA v123 Nr99 taxonomic database.

Statistical AnalysisAlpha and beta diversity metrics will be compared between vaginal and caesarean-born infants at different time points using non-parametric tests (e.g., Kruskal-Wallis, Mann-Whitney U). Differential abundance analysis will be performed using appropriate statistical tests (e.g., DESeq2)